A direct structure determination using lectin-carbohydrate complex
| Project/Area Number |
15K12750
|
|---|
| Research Category |
Grant-in-Aid for Challenging Exploratory Research
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Chemical biology
|
|---|
| Research Institution | Hokkaido University |
|---|
Principal Investigator |
Sakai Ryuichi 北海道大学, 水産科学研究院, 教授 (20265721)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
及川 雅人 横浜市立大学, 生命ナノシステム科学研究科(八景キャンパス), 教授 (70273571)
|
|---|
| Research Collaborator |
TANAKA Yoshikazu 北海道大学, 大学院・先端生命科学研究院, 准教授 (20374225)
|
|---|
| Project Period (FY) |
2015-04-01 – 2017-03-31
|
| Project Status |
Completed (Fiscal Year 2016)
|
| Budget Amount *help |
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
|
|---|
| Keywords | レクチン / X-線構造解析 / 直接構造決定 / 結晶構造解析 / 糖プローブ / 低分子プローブ / 構造決定 |
|---|
| Outline of Final Research Achievements |
Lectins, carbohydrate binding proteins, bind to sugars. We have isolated a galactose binding lectin CchG from the marine sponge Cinachyrella sp. that forms a rigid tetramer in the crystalline state. If the lectin-carbohydrate complex can be crystallized and its structure solved, the structure of the carbohydrate bind to the protein will be visualized. We extended this idea to determine the structure of glycosylated small molecules; that is, if a small molecule was glycosylated with the sugar that can bind to the lectin, the structure of the glycoside would be visualized directly by X-ray crystallography at the atomic level.
We thus tested our concept by synthesizing 1-fluoro-2,4-dinitrobenzene-5-(1-aminolacoside) (LacDNFB) which can readily couple with amino acids. In crystallization of CchG-lactose, CchG-LacDNFB, and CchG-LacDNFB-alanine complexes, and crystal structure for CchG-lactose showing that +1-end of the sugar had a space to hold molecule to be visualized, was solved.
|
Report
(3 results)
Research Products
(1 results)
